103 results about "Rice black-streaked dwarf virus" patented technology

The invention provides a method for rapidly identifying the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, belonging to the field of plant protection. Three primers are designed according to the nucleotide sequence of the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, which are respectively [A] RB1_S9_F, [B] RB2_S9_F and [C] RB_S9_R, wherein the combination of A and C detects the rice black-streaked dwarf virus and the combination of B and C detects the southern rice black-streaked dwarf virus. After the total RNA is extracted fromthe samples, the two viruses of the rice black-streaked dwarf virus and the southern rice black-streaked dwarf virus can be detected by only one RT-PCR, thus realizing the identification of two viruses. Compared with the traditional method in which one RT-PCR can only detect one virus, the invention greatly reduces cost, reduces time, and is a detection method which is special, sensitive, economic and convenient.

The invention provides a pesticide composition, and in particular relates to an active pesticide composition, compounding one or more of toxic fluoride phosphate, the weight ratio of toxic fluoride phosphate to other active substances is 30:0.15 to 3.3:30, and the content of effective components by weight is 1%-90%. The compound composition can be processed into water dispersible powder, suspending agent, microemulsion, aqueous emulsion, water dispersible granule, dry suspension emulsion and seed coating agent and the like, after accessory ingredient or carrying agent is added. The composition can be used to prevent and treat virus diseases caused by rice stripe virus, rice dwarf virus, maze rough dwarf virus, rice black-streaked dwarf virus, tomato virus, cucumber mosaic virus and tobacco mosaic virus, and has the beneficial effects of enhanced effect, concurrent treatment, postponed pesticide resistance, decreased dosage of toxic fluoride phosphate and reduced cost.

The invention provides an anti-rice black-streaked dwarf virus field inoculation identification method for rice varieties, and belongs to the technical field of agricultural science. A piece of rice field with certain conditions is selected as an identification nursery, different rice varieties are sown in special stages, it is identified that virus transmission interbody small brown rice planthoppers in the nursery have certain virus carried rate and population density after wheat harvest, and after rice grows to a specific growth period, the anti-rice black-streaked dwarf virus level of the rice varieties is evaluated according to the severity degree of disease. With the method, anti-rice black-streaked dwarf virus identification can be performed on a large number of rice varieties at a time, and the defects that the manual identification scale in a laboratory and the identification variety number are small are overcome. The identification method can be applied to the multiple fields of screening of anti-rice black-streaked dwarf virus rice varieties, variety resistance evaluation, resistant variety breeding, researching on the law of genetics of resistance and the like.

The invention discloses a method for obtaining sogatella furcifera carrying SRBSDV (southern rice black-streaked dwarf virus) and an application sogatella furcifera. The method for obtaining the sogatella furcifera carrying the SRBSDV, comprises the following steps of: feeding the sogatella furcifera by utilizing rice infecting the SRBSDV or cryopreserved rice diseased plant so that the sogatella furcifera obtains the SRBSDV, transferring the infected sogatella furcifera to a healthy rice to be fed till passing through a circulation period of virus, and detecting the sogatella furcifera by utilizing an RT-PCR (reverse transcription-polymerase chain reaction) technology and a serology method to obtain the sogatella furcifera carrying the SRBSDV, wherein the carrier rate of the sogatella furcifera can reach more than 50% through a manual feeding manner based on the experiment. Through utilizing the method for obtaining the sogatella furcifera carrying the SRBSDV, the obtained sogatella furcifera carrying the SRBSDV can be applied to screening of SRBSDV-resisting monoclonal antibody to carry out a rice inoculation experiment to identify the resistance of the rice, screen disease-resistant variety and provide a service of the breeding for disease resistance. The invention can be further applied to researching mutual relation between the SRBSDV and a vector insect sogatella furcifera and provides powerful theoretical evidence for prevention and treatment on the viruses.

The invention discloses an immunodetection kit specially used for rice black-streaked dwarf virus in single planthopper and a using method thereof. Specific polyclonal antibodies of the rice black-streaked dwarf virus (RBSDV) are utilized to build a dot immuno-binding assay (DIBA) detecting method which is used for building the detection kit for rice black-streaked dwarf virus, main reagents of the kit are working solution or concentrated solution, and the kit has the characteristics of convenient use, simple operation, and the like. And simultaneously, the use of the rice black-streaked dwarf virus detection kit can be a standard program or a reduced program, which can greatly broaden the using ranges and applicable persons of the kit and facilitate the population and application of the technique to establishment units, and further, the kit is applied in the fields of plant disease prognosis and prediction and the formulation of prevention and curing strategies, and the like. In addition, the kit can be used for measuring the inoculation strength in varietal resistance seed selection of rice black-streaked dwarf virus, and the like.

The invention relates to an RT-PCR detection method of an ordinary rice black streaked dwarf virus (RBSDV). A pair of used primers are obtained by the method that the first section of RNA of an RBSDV is specific, and nucleotide sequences of the first section of RNA of the RBSDV and the first section of RNA of similar viruses are researched and downloaded from NCBI by utilizing the conservative property of the evolution of viral nucleotide sequences; and the sequences are compared by using MEGA4.0 software to find out a primer candidate area which is reported by different researchers and shared by the RBSDV nucleotide sequences and is different from other viral sequences from a mas file. Proved by BLAST results, the sequences which can 100 percent cover the primer 1 and the primer 2 and 100 percent same as the primer 1 and the primer 2 are all the RBSDV nucleotide sequences. RBSDV and SRBSDV viruses can be exactly separated by using the pair of primers for carrying out RT-PCR detections on the RBSDV and SRBSDV viruses.

The invention discloses molecular markers closely linked to an RBSDV (rice black-streaked dwarf virus) major QTL (quantitative trait locus). The essence of the invention is that according to the linkage and segregation rules, the F2 population of the hybrid generation of Tetep and RBSDV susceptible varieties is taken as the mapping population and the SSR (simple sequence repeat) markers RM5626 and RM7097 on the No.3 chromosome, which are closely linked to the RBSDV resistant major QTL are obtained through analysis of linkage between the results of evaluation of disease resistance and the SSR marker data. Whether a plant has resistance to RBSDV can be judged by detecting the SSR marker banding pattern of the single rice plant of the hybrid combination generation of Tetep and derived varieties (lines) thereof and the RBSDV susceptible varieties. After the SSR markers are applied to assistant selection breeding of the RBSDV resistant varieties of Tetep and derived varieties (lines) thereof/RBSDV susceptible varieties, the defects of poor stability and repeatability of RBSDV resistance evaluation, time and labor waste, high technical threshold and the like in conventional breeding can be overcome and the results can conduce to simplifying the selection method and improving the breeding efficiency and then accelerating the breeding process of RBSDV resistant varieties.

The invention relates to RNA (Ribonucleic Acid) interference carriers of RSV (Rice Stripe Virus) and RBSDV (Rice Black-Streaked Dwarf Virus) as well as a construction method and application thereof, belonging to the technical field of biology. The invention establishes RNA interference carriers pMCG161+/-D and pCMBIA1301+/-D of the RSV by using sequences shown as SEQ ID NO1. The invention provides an interference virus gene expression carrier for plant disease-resistant gene engineering as well as a construction method and application thereof and provides a new concept for the application of a transgenic technology and RNA interference in biological resistant breeding. By transplanting the carrier to rice, a virus-resistant plant is successfully acquired, the expected target of obtaining a RSV and RBSDV-resistant rice new material according to the principle of RNA interference is realized, a successful example is provided for plant resistant breeding and gene engineering of RNA interference mediums and the blank of researches on the RSV and RBSDV resistance of the RNA interference mediums in the field is made up.

The invention discloses a preparation method and biological activity of a pentadiene ketone compound, containing 1,3,4-oxadiazole sulfo-ethyoxyl, with a function of preventing and treating a plant virus disease, namely a compound expressed in a general formula (I) and a preparation method of the compound. According to the method, by using substituted carboxylic acid, hydrazine hydrate, carbon disulfide, acetone, 4-substituted hydroxy benzaldehyde, substituted aromatic aldehyde, 1,2-dibromoethane and the like as raw materials, methanol, ethanol and N,N-dimethylformamide as solvents and potassium carbonate, sodium hydroxide and potassium hydroxide as catalysts, pentadiene ketone compound containing 1,3,4-oxadiazole sulfo-ethyoxyl is prepared through seven-step synthesis. The compounds I3, I4, I6, I11, I13, I14, I16 and I17 have good functions in cucumber mosaic virus, tobacco mosaic virus, southern rice black-streaked dwarf virus and the like. The formula is shown in the specification.

The invention provides a rapid detection technology for rice black-streaked dwarf viruses in plants such as rice, wheat and corn and the like and an application method thereof in the diagnosis of diseases. By the method, the interference of southern rice black-streaked dwarf disease can be eliminated, and plant samples of the rice black-streaked dwarf viruses can be rapidly identified, so that targeted preventive strategies can be used, and the loss brought by the diseases is reduced.

Preparation and application of bispyrazole carboxamide derivative to the control of rice black streaked dwarf disease. The invention discloses a bispyrazole carboxamide compound, which is synthesized by using carboxamide structure as the core, and introducing bispyrazole ring. The invention also discloses the preparation method of the bispyrazole carboxamide compound and application of the compound to the control of rice black streaked dwarf disease. The invention has the advantages that through the preliminary bioassay, the synthesized target compound has good activity on rice black streaked dwarf virus.

The invention relates to a rice black-streaked dwarf virus (RBSDV) resistant locus qRBSDV11 of a rice variety 9194 and a molecular marker method thereof. Four RBSDV resistant genetic loci of the RBSDV resistant variety 9194 are obtained by carrying out genetic linkage analysis on the genotype of a single plant F2 obtained by hybridizing the RBSDV resistant rice variety 9194 (male) with a susceptible variety Suyunuo (female) and the RBSDV resistance grade of a corresponding F2:3 family, wherein qRBSDV11 is between a marker RM26062 and a marker RM536. The RBSDV resistance levels of the resistant variety 9194 and derived varieties (lines) thereof can be predicated, and the selection efficiency of the RBSDV resistant rice can be greatly improved by detecting whether the resistant variety 9194 and the derived varieties (lines) thereof contain the RBSDV resistance genetic loci via the molecular markers of the RBSDV resistant genes.

The invention provides a method for SRBSV (southern rice black-streaked dwarf virus) detection on rice plants and accurate quantification of RNA (ribose nucleic acid) copy number. The method can be used for eliminating the interference of the SRBSV, the copy number of the virus RNA in the SRBSV plants is fast obtained, and further, the basis is laid for studying the SRBSV pathogenic mechanism, the molecular biology study and the like.

The invention discloses combinations of reference genes for gene function analysis on an RBSDV (rice black-streaked dwarf virus) infected plant. The combinations are obtained according to the following steps: analyzing the expression stability of a fluorescent quantitative PCR candidate reference gene in the RBSDV infected plant by utilizing geNorm and NormFinder, then the paired difference value V of a normalized factor is calculated after a new reference gene is introduced according to a geNorm program, and determining optimal reference gene combinations according to the variation of V, so as to obtain the optimal reference gene combinations for gene function analysis on the gene function analysis, wherein the optimal reference gene combinations are UBC and Actinl, and when the RBSDV infected plant is taken as a target, the optimal reference gene combinations (UBC and Actinl) can be used for obtaining more accurate and reliable gene expression results as compared with a single reference gene. The combinations can be used for researching the expression level and function of related genes of the RBSDV infected plant and the infecting mode of RBSDV.

The invention discloses a hybridoma cell strain excreting a monoclonal antibody (MAb) resisting a rice blackstreaked dwarf virus (RBSDV) and the application of the MAb. A coat protein (CP) of the RBSDV is expressed into antigen immune BALB/c mouse by a prokaryotic expression method, and the hybridoma cell strain 5G1 which can stably passage and excrete the MAb resisting the RBSDV is obtained through cell fusion, selection and cloning, wherein the preserving number is CGMCC No. 5537. Ascitic indirect enzyme-linked immuno sorbent assay (ELISA) valence of the 5G1 MAb is over 10<-6>, and the antibody type and the subtype are IgG1 and kappa chain. The antibody excreted by the cell strain and the coat protein of the RBSDV have specific immunobinding reaction. By the dot-ELISA detection method for detecting rice planthopper and the RBSDV in rice and established by using the 5G1 MAb as a core, the virus can be still detected when a single-head small brown rice planthopper is diluted to 1,600 microlitres and sick leaves are diluted (w/v, g/mL) in the proportion of 1:160. The preparation of the MAb resisting the RBSDV and the establishment of the detection method thereof provide technical and material support for the diagnosis, forecast and scientific prevention and control of rice viruses.

The invention discloses a method for quickly detecting southern rice black-streaked dwarf virus based on RPA. A primer pair is composed of two primers shown in the sequence 1 and the sequence 2 in a sequence list. The invention protects the method for detecting the southern rice black-streaked dwarf virus by using the primers. The method does not depend on precise test instruments, the operation method is simple, the primers can effectively amplify the target gene, and the method is high in specificity and higher in sensitivity; early quick detection of the virus can be achieved, and the method has an important significance for preventing and controlling occurrence and prevalence of the virus.

The invention relates to a rice variety IR36 black-streaked dwarf virus resistance site qRBSDV-1 and a molecular marking method and application thereof. According to the invention, a recombinant inbred line population constructed by crossing a rice black-streaked dwarf virus resistance variety IR36 and a black-streaked dwarf virus susceptible variety L5494 is taken as a positioning population, andthe black-streaked dwarf virus resistance identification is carried out by a field natural identification method, a resistance gene site, named qRBSDV-1, from a black-streaked dwarf virus resistancevariety IR36 is obtained, located between the markers AP-39.6 and RM104. The method verifies qRBSDV-1 by performing molecular detection and identification of the incidence of black-streaked dwarf virus by using a substitution line (No. N9) containing 93-11 introduced fragments only in the chromosome segment where qRBSDV-1 is located and parents in a Japanese sunny background. According to the invention, the resistance of the plant to the rice black-streaked dwarf virus can be predicted by detecting the bands at the molecular markers of AP-39.6 and RM104, and the breeding efficiency of the ricevariety resistant to the black-streaked dwarf virus is improved.

The invention discloses an osa-miR396d responding to rice black streaked dwarf virus infection, and application thereof. The invention further discloses application of RNA (Ribonucleic Acid) shown in sequence 1 in suppression of gene expression of growth regulators; the growth regulators are shown in the sequence 2 of a sequence table. Expectedly, antiviral plants can be obtained by using the osa-miR396d gene of rice, which is beneficial to agricultural production.

The invention relates to an RT-PCR detection method of a south rice black-streaked dwarf virus (SRBSDV). In the detection method, SRBSDV specific primers are used for carrying out RT-PCR amplification on a sample to be detected and then carrying out electrophoretic identification. The specific primers are obtained by the method that by utilizing the conservatism of the evolution of viral nucleotide sequences, the tenth section of RNA of SRBSDV and nucleotide sequences of the tenth section of RNA of similar viruses are searched and downloaded from NCBI/GenBank, the sequences are compared by utilizing MEGA4.0 software, and then a pair of primers which are shared by all the SRBSDV nucleotide sequences reported by different researchers and are different from other viral sequences are found out from a file with an extension name of mas. Proved by a BLAST result, sequences which 100 percent cover the primers and 100 percent same as the primers are all the SRBSDV nucleotide sequences. The primers can be used for accurately separating SRBSDV diseased plants from ordinary RBSDV diseased plants without a nested RT-PCR detection.

The invention discloses a primer and a reagent kit capable of being used for detecting southern rice black-streaked dwarf virus (SRBSDV) and also discloses a method for quantitatively detecting the SRBSDV by the reagent kit. According to the method, firstly, the primer is synthesized; then, an RNA (ribonucleic acid) extraction reagent kit is utilized for extracting RNA of the primer from samples; the RNA is used as a template, in addition, a cDNA (complementary deoxyribonucleic acid) synthetic reaction system is utilized for synthesizing cDNA, then, positive standard samples are prepared and are subjected to real-time fluorescence quantification PCR (polymerase chain reaction) procedure drawing, and a standard curve of circulation number-initial concentration logarithmic values with fluorescence signals reaching the threshold values is obtained; and finally, a real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) system is prepared by using the synthesized cDNA as the template, the amplified reaction is carried out through the real-time fluorescence quantification RT-PCR procedure, and the initial concentration value of the SRBSDV in the samples is measured according to the collected fluorescence signals and the standard curve. The method has the advantages that the precision is high, the stability is good, the repeatability is good, the specificity is high, the sensitivity is high, and the like.

The invention discloses a monoclonal antibody hybridoma cell line for secreting southern rice black-streaked dwarf resistant viruses and monoclonal antibody application thereof. Polypeptide of 12 amino acids at the C end of a capsid protein of southern rice black-streaked dwarf viruses (SRBSDVs) coupled with bovine serum albumin is used as an antigen to immune a BALB/c mouse, one hybridoma cell line 3F1 capable of performing stable passage and secreting an SRBSDV monoclonal antibody is obtained through cell fusion, screening and cloning, and the preservation number of the hybridoma cell line 3F1 is China general microbiological culture collection center number (CGMCCNo.) 5535. Monoclonal antibody ascites indirect enzyme-linked immunosorbent assay (ELISA) valence of the 3F1 reaches above 10-6, an antibody type and a subclass are immunoglobulin G (IgG1) and a kappa chain, the monoclonal antibody of the hybridoma cell line has specific reaction with a capsid protein subunit of SRBSDV56kD, and a method for detecting rice planthoppers and dot-ELISA of SRBSDV in rice is built by using the monoclonal antibody of the 3F1. When a single-head white backed rice planthopper is diluted to 6400mul, the viruses can still be detected when diseased leaves are diluted according to 1:320 times (w/v, g/mL). The preparation of the SRBSDV resistant monoclonal antibody and the building of the detection method provide technology and material supports for diagnosis, prediction and scientific prevention and control of the rice viruses.

The invention provides a rapid detection technology for rice black-streaked dwarf viruses in plant hopper and a method for applying the rapid detection technology to disease diagnosis and prediction. By the method, the interference of southern rice black-streaked dwarf viruses can be eliminated, and whether the rice black-streaked dwarf viruses are carried in the plant hopper is rapidly identified, so that a pertinent control strategy is adopted to reduce the loss caused by diseases.

The invention provides RT-PCR used for detecting rice black streaked dwarf virus (RBSDV) and southern rice black-streaked dwarf virus (SRBSDV) and an application of the RT-PCR and belongs to the technical field of PCR detection. Through comparison of genomic sequences of RBSDV with SRBSDV, a conservative-area target sequence is determined, a specific primer pair used for detecting the RBSDV and the SRBSDV is designed, and the nucleotide sequences are as shown in SEQ ID NO. 1-4. The invention further provides a method for detecting the RBSDV and the SRBSDV and a detection kit. The method has the advantages of accurate detection, high flexibility, high specificity, convenience and rapidness, crop infection of the RBSDV or the SRBSDV can be distinguished, low cost is achieved, and good economic value and application prospect are realized.